Prognostic role of clinical presentation, cytological picture and response to treatment in canine centroblastic lymphoma.

Centroblastic lymphoma (CBL) is the most common type of lymphoma in dogs and it usually responds well to chemotherapy. The aim of the study was to provide useful prognostic factors for dogs with CBL. Data regarding sex, breed, age, signalment, treatment and clinical course of the disease from 52 dogs diagnosed with centroblastic lymphoma (CBL) with cytology and immunocytochemistry were provisionally collected and related to the treatment outcome and survival. More than 80% of dogs were treated with chemotherapy and achieved complete remission in 80% of cases. Among the prognostic factors positively related to the overall survival time of dogs with CBL were: the application of chemotherapy, achieving a complete remission, application of at least one additional chemotherapeutic agent to the basic protocol, especially the administration of mitoxantrone and asparaginase. Moreover, mitotic count 14 or higher measured in cytological slides in the area of 2,37 mm2 have been linked to shorter overall survival in dogs with CBL.

Malignant thymoma - the most common neoplasm in goats.

In the literature the occurrence of thymomas in goats varies from 0.7 to 25%, depending on the study. Therefore the current investigation was carried out to determine the prevalence of thymoma in goats in Poland. Between 2007 and 2018 at the Warsaw Veterinary Faculty 399 goat autopsies and ultrasound examinations of the chest in other 145 goats were performed. Mediastinal tumors were diagnosed during post mortem examination in 2 goats. Additionally, ultrasound examination of the chest revealed a large mass close to the heart in the thoracic cavity in 1 case. This goat was euthanized and an autopsy confirmed a mediastinal tumor. Histopathological examination, with immunohistochemical tests to anti cytokeratin, p63 and p40 confirmed thymomas in all three cases. In our study thymomas were found in 0.5% (95% CI: 0.1% to 1.8%) of examined goats and they represented the most common malignancy in this species.

The animal-dependent risk factors in canine osteosarcomas.

Canine osteosarcoma (OSA) is a malignant neoplastic tumor, which develops from the primitive mesenchymal stem cell, that has or can acquire the capacity to produce neoplastic osteoid with possible neoplastic bone formation. Predisposition of some dog breeds to OSA indicates genetic background of oncogenesis. The aim of the study was to characterize animal-dependent risk factors for canine osteosarcoma development in Poland. The study was conducted on canine patients diagnosed cytologically or histopathologically as having OSA, and data on age, breed, sex, as well as tumor location and character were recorded. No sex predisposition to OSA was observed, mongrels were significantly underrepresented. Large and giant dogs accounted for 47% and 35% of all pedigree dogs, respectively, and both proved predisposed to OSA. A vast majority of OSA developed in the skeleton (appendicular skeleton was more commonly affected than axial skeleton), soft tissues were affected less often. Rottweiler dogs are strongly predisposed to OSA, suggesting that the genetic background is involved in the tumor development, and indicates that dogs of this breed are a promising object for further studies on OSA pathogenesis.

Metastatic lymphadenomegaly in dogs - cytological study.

Enlarged lymph nodes are frequently examined cytologically in dogs and metastatic lymphadenomegaly of various origin is a common cytological finding in these cases. In this study we aimed to examine epidemiological data, and to determine factors affecting the location of neoplastic metastases in the lymph nodes. Samples for cytological examination were obtained by fine-needle biopsy (FNB) of enlarged lymph nodes and stained with Giemsa solution. Cases meeting the following criteria were enrolled in the study: lymphadenomegaly detected in clinical examination, presence of primary mass confirmed by cytopathology or histopathology as a solid malignant tumour, and cytological diagnosis of metastatic tumour. Cytological pattern of lymph node involvement was classified as low, medium and massive. During study period 125 dogs met the eligibility criteria, with age ranged from 1.8 to 19 years. No sex predisposition to particular types of tumors was observed, except for adenocarcinoma which was diagnosed in females more often. Metastatic tumors were various in origin, with predominance of mast cell tumors, adenocarcinomas, and melanomas. Massive involvement predominated in all lymph nodes affected. Neoplastic lymphadenomegally is recognized usually in older dogs, with female predisposition related to dissemination of mammary cancers. Mast cell tumor, adenocarcinoma and melanoma are the most common causes of metastatic lymphadenomegaly, and in the vast majority of the cases massive lymph node involvement is observed.

The animal-dependent risk factors in canine T-cell lymphomas.

Malignant lymphomas are one of the most common malignancies occurring in dogs; among them T-cell tumours are less commonly recognized. Recently, many authors have recommended cytology as a sufficient diagnostic method for canine lymphomas, especially if supported by immunocytochemistry or flow cytometry. The aim of the study was to characterize animal-dependent risk factors in canine T-cell lymphomas (TCLs) in Poland, including specific cytological subtypes. Determination of the type and subtype of the tumour was made based on the updated Kiel cytological classification adopted for dogs as previously described. Two breeds turned out predisposed to TCL (dog de Bordeaux and Boxer) while no predisposition to B-cell lymphoma could be evidenced. Dogs with low-grade lymphoma were significantly older than those with high-grade lymphoma.

Factors affecting the diagnostic utility of canine and feline cytological samples.

OBJECTIVES: Cytology is a quick, inexpensive, minimally invasive and widely available diagnostic method, but rigorous cooperation between clinical cytopathologist and physician is necessary to obtain clinically useful results. The aim of this study was to identify factors that affect clinical usefulness of a cytological result when the cellular material was collected by a general veterinary practitioner and examined by a clinical cytopathologist. MATERIALS AND METHODS: Analysis of 100 fine-needle aspirates performed by private veterinarians and examined by the clinical cytopathologist. Factors dependent on a general veterinary practitioner who performed the biopsy such as comprehensiveness of a cover letter, number of smears sent and macroscopic appearance of smears, were included in the analysis. Patient species and location of the lesion were also included. RESULTS: Only two factors turned out to favour a cytological diagnosis: good macroscopic appearance of smears and superficial location of the lesion. Nevertheless, inclusion of the medical history of a patient in a cover letter proved to help a clinical cytopathologist make clinically useful suggestions. CLINICAL SIGNIFICANCE: Preparation of good quality smears and provision of a comprehensive cover letter will increase the likelihood of obtaining clinically useful cytological reports.

Cytodiagnostics of canine lymphomas - possibilities and limitations.

Malignant lymphomas are one of the most common malignant tumours occurring in dogs. The basic method of lymphoma diagnosis in human, as well as in canine oncology is histopathology supported by immunohistochemistry. It was suggested that in veterinary medicine excisional biopsy of lymph node and histopathology should be considered only where the cytologic diagnosis is equivocal or needs to be confirmed. There are at least three basic reasons for which cytological examination ought to be accepted as a sufficient and reliable diagnostic method for lymphoma in dogs. Firstly, most dog owners consider the fine-needle biopsy as an acceptable non-harmful method of sample collection. Secondly, an increasing number of studies recommend cytology as an accurate test for diagnosing and subtyping canine lymphoma. Finally, the vast majority of canine lymphoma subtypes belong to 4-5 categories characterized by a typical cytological picture. Immunocytochemical staining of cytological smears gives new diagnostic possibilities, such as detection of markers better characterizing given growth or a potential goal for target therapy in individual cases (for example inhibitors of platelet-derived growth factor).

Cytomorphometry of serosal effusion in dogs.

Cytomorphometry made on cytological slides is the quantitative method of precise analysis of cellular structures, including both cytoplasm and nucleus. The aim of this study was to describe cytomorphometric parameters of mesothelial cells in the course of benign reactive and malignant proliferation and to compare them to carcinomas and adenocarcinomas located within serosal cavities in dogs. The second aim was to evaluate applicability of cytomorphometry to diagnostics of diseases causing accumulation of effusion in serosal cavities. Cytological samples of normal and non-malignant mesothelium, mesothelioma and various carcinomas were collected from dogs. Cytomorphometry was made on the smears stained with Giemsa solution. Mean nuclear and cellular perimeter, mean nuclear and cellular area, mean nuclear and cellular diameter, and mean nuclear and cellular roundness were determined. Moreover, nuclear to cytoplasmic ratio (N/C) was calculated. The data revealed statistically significant differences for all parameters, excluding mean nuclear perimeter, between compared groups. Normal mesothelium cells and their nuclei were significantly smaller and more elongated than cells and nuclei of both benign reactive and malignant neoplastic mesothelium. Only a few differences were observed between benign reactive mesothelium cells and mesothelioma cells - mean nuclear area and mean nuclear diameter of benign reactive mesothelium cells were significantly larger and N/C ratio was higher in comparison to mesothelioma cells. Even though some significant differences were observed, considerable overlap of these cytomorphometric parameters in animals with different diseases limited practical role of these observations. Cytomorphometric analysis of cellular samples collected from dogs with proliferative processes affecting serosal cavities can be only an auxiliary method increasing accuracy of preoperative diagnosis.

Epidemiological and morphological analysis of feline injection site sarcomas.

Feline injection site sarcomas (FISS) are malignant neoplasms of mesenchymal origin which arise in sites of injections in cats. The prevalence is estimated between 1 in 1000 and 1 in 10 000 vaccinations in the United Kingdom. The aim of this study was to estimate the incidence of FISS in Poland and to analyse clinical aspects and histological and cytological features of injection site sarcomas. In our study the prevalence of FISS was 0.16% (16 FISS on 10.000 of cats) in feline patients in one of a veterinary surgery which conducts the general practice and 85 on 10.000 cats in a practice focused on veterinary oncology. The most typical microscopic features of FISS found in the present analysis were: the presence of perilesional scarring and inflammation, aggregates of lymphocytes at the tumour periphery, moderate but usually marked cellular pleomorphism and intralesional necrosis. The most typical cytologic features of FISS found in present study were: the presence of neutrophils, marked cellular pleomorphism, the presence of lymphocytes and macrophages, the presence of extremely large nuclei in the neoplastic cells, and high sample cellularity.

Large granular lymphoma in six cats.

Large granular lymphomas (LGLs) comprise a specific group of lymphomas regardless of classifi- cation scheme. An LGL consists of cells that show less or more mature morphology, but typically neoplastic cells possess cytoplasmic azurophilic granules clearly visible during cytological examin- ation. The aim of the present study was to present clinical and cytological data on large granular lymphomas in cats and to analyses the therapeutic responses in treated cases. During the period from 2012 to 2014 six cats were as having large granular lymphoma. In one cat a nasal form of LGL was recognized, a systemic form was recognized in another cat, and in four cases an alimentary form was recognized. Cellular samples for cytopathology were collected from the cat with nasal cavity mass, from the enlarged mandibular lymph node and thoracic cavity from second cat, and in four cats from the abdominal mass during ultrasound-assisted fine-needle biopsy. Therapy was introduced in 5 of the 6 cats. In two cases palliative therapy with glucocorticoids was conducted, in two cases chemotherapy with COP protocol, and therapy with masitinib in one case. The median of survival time for cats treated with anticancer therapy was 9 months, the median of survival time for cats treated with glucocorticoids was 1.5 months. In conclusion, large granular lymphomas, especially the alimentary form, are a relatively common type of lymphoma in cats. Simple diagnostic methods such as clinical examination, imaging techniques and routine cytology are sufficient in majority of cases. Despite aggressive behavior and poor general prognosis, conventional chemotherapy lead to a good response in some treated cats regardless of anatomic form and histologic grade of malignancy.

Fungal rhinitis in dogs.

Fungal rhinitis and sinusitis in dogs are quite common reasons of chronic nasal discharge and rhinoscopy in such cases is commonly suggested. Forty three dogs were examined using rhinoscopy because of the presence of chronic airway symptoms. Clinical examination, routine hematology and serum biochemistry profiles, nasal and frontal sinus radiographs were made in all animals. Additionally, computed tomography in one dog was performed. Samples for histopathology were taken from 9 patients during rhinoscopy, additionally, from 8 of these patients samples for cytopathology were collected by blind nasal swab technique. In 9 of 43 dogs (20,5%), all males aged 1 to 13 years, examinations led to a diagnosis of fungal rhinitis. In 2 cases a diagnosis of fungal rhinitis was obtained based solely on cytopathology, while in 7 cases - mycosis of nasal mucosa was confirmed by histopathology. The present study revealed that cytopathological examination of nasal swabs has a low diagnostic value in the case of nasal infections in dogs. Although, in some dogs cytopathology, together with other widely available diagnostic techniques was sufficient to reliably diagnose fungal rhinitis, histopathology of samples collected during rhinoscopy is still the gold standard in such cases.

Using of immunocytochemistry in differential diagnosis of neoplasms of serosal cavities in dogs.

The presence of tumor within the serosal cavities, often connected with accumulation of serosal effusion, is a quite common problem in the small animal veterinary medicine. The first step in diagnosis of such cases is cytopathological examination. The aim of the present study was to evaluate the usefulness of cytology and immunocytochemistry, using commercially available antibodies (anti-cytokeratin, anti-vimentin, and anti-desmin), in differential diagnosis of malignant tumors located within serosal cavities in dogs. The final cytological diagnosis of carcinoma/adenocarcinoma, sarcoma, and mesothelioma was obtained on the basis of routine cytopathology and immunocytochemistry, and then confirmed by histopathology and immunohistochemistry. Cytoplasmic immunoreactivitiy of normal mesothelid cells and cytoplasmic immunoreactivity of hyperplastic mesothelial cells revealed constant and strong expression of all examined intermediate filaments: cytokeratin, vimentin and desmin. Application of routine cytopathology and immunocytochemistry allowed 32 neoplastic tumors to be detected: 19 cases of carcinomas/adenocarcinomas, 6 cases of sarcomas, 7 cases of mesotheliomas. Immunostaining of cytopathological samples with chosen set of antibodies: anti-cytokeratin, anti-vimentin, anti-desmin is a useful, and low invasive test for differentiation between mesotheliomas and carcinomas/adenocarcinomas in dogs.

High agreement of routine cytopathology and immunocytochemistry in canine lymphomas.

The aim of this study was to compare the concordance of immunophenotype established with routine cytopathology (slides stained with Giemsa solution) and immunocytochemistry according to CD3 and CD79 alpha immunoreactivity. The study was performed on cytological samples of 70 canine lymphomas subtyped on the basis of the updated Kiel classification system. Additionally, cytologic samples were examined immunocytochemically for the CD3 and CD79 alpha antigens presence and thus immunophenotype of neoplastic growth was confirmed. The cytopathological and immunocytochemical diagnoses were then compared; in order to measure the concordance between immunocytochemistry (IC) result and Giemsa stain result of the same sample the Cohen's kappa coefficient was calculated. On the basis of the results of immunocytochemistry of 70 cases of canine lymphoma examined 42 were recognized as B cell lymphoma and 28 as T cell lymphoma. Full accordance between the results of routine cytopathology and IC was obtained in 63 out of 70 examined dogs (90% of cases). It can be concluded that cytopathological examination of Giemsa stained smears is helpful in determining the lymphoma cells immunophenotype. Additionally, it seems that combination of routine cytopathology and immunocytochemistry in cases of canine lymphomas allows to obtain the precise diagnosis in 90% of cases, and allows to receive most important information that is necessary for planning of appropriate therapy and to determine prognosis. Finally, this routine procedure allowed to eliminate the need of collection of tissue samples during surgery or core-biopsy and thus time, cost and patient discomfort related to more complex and invasive medical procedures can be easily reduced.

Cytopathological diagnosis of visceral histiocytic sarcoma in five dogs.

Histiocytic sarcoma is a malignant neoplastic proliferation of atypical histiocytes with tendency to spread, characterized by fast progression to disseminated form--disseminated histiocytic sarcoma. Cytopathology is a low, invasive, cheap, and quick method of diagnosis commonly used in veterinary oncology. The aim of the presented study was description of cases of visceral histiocytic sarcomas in dogs diagnosed by cytopathology and immunocytochemistry. The study was conducted on 5 dogs which were brought to the veterinary clinic because of unspecific clinical signs and tumoral masses recognized in the thoracic or abdominal cavity. Samples of cells were collected during ultrasonography-assisted fine-needle aspiration biopsy (FNAB), smears were stained with Giemsa method and immunocytochemistry (CD3, CD79alpha, cytokeratin, vimentin, desmin) was also performed in all patients. Four of five dogs were Bernese mountain dogs, nonspecific clinical signs of systemic disease were present in all cases. Visceral mass or masses were detected by ultrasonography or radiography. Final diagnosis of histiocytic sarcoma was obtained on the basis of routine cytopathological examination and confirmed by immunocytochemistry. On the basis the results obtained it can be stated that in cases of typical clinical and cytopathologic pictures, examination of cellular samples collected during ultrasonography-assisted fine-needle biopsy supported by some immunocytopathological characteristics seems to be sufficient method of diagnosis of histiocytic sarcoma in dogs. Visceral histiocytic sarcoma should be included into differential diagnosis in every Bernese mountain dog with nonspecific clinical signs, ambiguous results of hematologic examination and when tumoral mass or masses within a body cavity were detected in imaging techniques.

Histopathology in diagnosis of broiler chicken and layer diseases--review of cases 1999-2010.

The aim of this study was to estimate the prevalence of histopathological lesions in the different organs in relation to the commercial-type and the age of birds (i.e. broiler chickens and layers). During the period 1999-2010 a total of 189 cases was submitted to the Division of Animal Pathomorphology, Department of Pathology and Veterinary Diagnostics at WULS. Most cases were found in broiler chickens (66.7%). The majority of the histopathological lesions were detected in the liver and lymphoid organs. In of 29% cases of hepatic injury pathognomonic lesions associated with inclusion body hepatitis (IBH) were found. The mean age of birds was 23 days. Among IBH cases proventriculitis (58%) was more often found than gizzard lesions (25.8%). Interestingly, we noted some intranuclear inclusions in the epithelial cells within the proventriculus. A low percentage of histopathological evidence of infectious bursal disease (IBD) was reported in chickens. The gastrointestinal tract was the second most frequent predilection site for histopathological lesions. Histopathological findings within the heart and lungs were less common and were more often seen in the upper respiratory tract. Cases of infectious laryngotracheitis (ILT) were registered in broiler chickens (3.2%, mean age 37 days) and in layers (4.8%; mean age 196 days). Lesions associated with Marek's disease, avian leukosis and fowl pox were recognized only in layers, respectively in 3.2% (mean age 176 days), 1.6% (mean age 205 days) and 1.1% (mean age 196 days) of all cases. Avian encephalomyelitis (AE) was noted only in 0.5% of all cases.

Mismatch between transcriptomic and histopathologic picture of canine lymphomas.

Lymphoma is one of the most common malignant tumours occurring in dogs. Since there is a constant need for new, more comprehensive laboratory diagnostic tools which permit the precise determination of many tumour-related factors we decided to verify whether the use of microarray analysis could be helpful in classifying lymphomas. The study was performed on samples collected from 7 dogs in which multicentric lymphoma was recognized. Among this group we were able to identify one sub-cluster of transcriptionally similar tumours, which completely differed in terms of the histopathological examination. Among them there were one diffuse large B cell lymphoma, one diffuse macronucleolated medium-sized cell lymphoma and one pleomorphic mixed small and large T-cell lymphoma. The lymphomas belonging to the sub-cluster differed from other analysed tumours in the expression of more than 100 genes of which only 18 were described earlier in regard to lymphomas.

The first description of gastric Helicobacter in free-ranging wild boar (Sus scrofa) from Poland.

Specimens of gastric mucosa of 17 free-ranging wild boars (Sus scrofa) shot in the Central Poland during 2007/2008 hunting season were investigated for the presence of Helicobacter species. Histopathology, Helicobacter genus-specific 16S rRNA PCR, and DNA sequence analysis were employed. In PCR analysis the presence of Helicobacter's DNA was detected in one stomach. Obtained sequence analysis showed its relatedness to Helicobacter heilmannii type 2. In histopathology of the PCR-positive sample the presence of tightly coiled spiral bacteria was detected on the surface of the antral mucosa, in gastric pits and lumen of the upper parts of antral glands. Potential pathologic significance of the presence of Helicobacter in the stomach of free-ranging wild boars was obscured by the parasitic invasion-caused gastritis, and remains unknown.

Several cases of dirofilariosis accidentally diagnosed in dogs from Poland, including two PCR positive Dirofilaria repens cases.

The aim of the present study was a description of several microfilaremia cases diagnosed in dogs from Poland. The study was conducted on cytological samples of pathologic masses and peripheral blood smears taken for diagnostic purposes from dogs showing a various clinical findings. The presence of the parasites in tissue samples was observed in 8 dogs, additionally, in 2 of these dogs PCR analysis of blood samples revealed the presence of D. repens DNA.

Cytopathology of canine lymphomas (100 cases).

Malignant lymphoma is one of the most common malignant tumours occurring in dogs. Fine needle aspiration biopsy (FNAB) is an excellent, specific diagnostic procedure used to assess pathological processes in lymph nodes. The aim of the present study was to conduct a cytopathological analysis of lymphoma in dogs and to analyse some epidemic aspects of occurrence of lymphoma in 100 dogs using Giemsa stained slides. Samples were obtained by fine needle aspiration biopsy, fine needle non-aspiration biopsy, lymph node impression smears and by examination of body cavity effusions. The determination of the type and subtype of tumour was made on the basis of the updated Kiel classification adopted for dogs. Based on cytopathological analysis, the lymphoma was diagnosed in 100 dogs: 44 were female and 56 male. The animals' age ranged from 1.5 year to 15 years (median: 7.5 years), the animals were of different breeds (72% of all dogs belonged to 28 different breeds) and crossbreeds (28%). In 29% of dogs the regional or general lymphadenomegaly was the only clinical sign observed, in remaining cases (71%) at least one abnormality connected to lymphoma was found. Among all diagnosed lymphomas, high-grade lymphomas were more prevalent (86% of all cases) than low-grade lymphomas (14% of all cases). The possibility of boxers having a predisposition to T cell lymphoma development could be also suspected.

Practical aspects of immunocytochemistry in canine lymphomas.

The aim of this study was to evaluate the utility of immunocytochemistry in a standard veterinary practice and to determine the immunophenotype of tumor cells in cases of multicentric lymphoma in dogs by immunocytochemical analysis of fine-needle biopsy specimens. The study was performed on cytological samples collected from 54 dogs, in which multicentric lymphoma was recognised based on clinical data, cytology or cytology and histology, and follow-up information. Diagnosis of lymphoma was established according to the updated Kiel classification. Immunocytochemical assays were conducted using commercially available antibodies to the pan T-lymphocyte marker CD3 and B cell antigen receptor complex CD79 alpha. Among all animals examined B cell lymphoma was recognized in 42/54 (77.8%) of cases, while in the remaining 12/54 (22.2%) of dogs T cell lymphoma was recognized. In 11 animals with lymphoma recognized cytologically, in which an entire lymph node was obtained for histology, the results of routine cytology and immunocytochemistry fully corresponded with findings revealed by histology and immunohistochemistry. Immunocytochemistry can be successfully conducted in smears stored at room temperature for 24 hours without changes of staining results. It can be stated that application of standard cytopathological assessment in connection with immunocytochemistry of lymph nodes samples collected from dogs with lymphoma is a method of choice for establishing final diagnosis, and avoids the need for reexamination or collection of tissue samples for histopathology and immunohistochemistry during surgical procedures in ambiguous cases.